An Intracellular Ammonium Transporter Is Necessary for Replication, Differentiation, and Resistance to Starvation and Osmotic Stress in <named-content content-type="genus-species">Trypanosoma cruzi</named-content>

ABSTRACT Trypanosoma cruzi, the etiologic agent of Chagas disease, undergoes drastic metabolic changes when it transits between a vector and mammalian hosts. Amino acid catabolism leads to the production of ammonium (NH4+), which needs to be detoxified. However, T. cruzi does not possess a urea cycl...

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Autores principales:	Teresa Cruz-Bustos, Evgeniy Potapenko, Melissa Storey, Roberto Docampo
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	Trypanosoma cruzi amino acids ammonia ammonium autophagy lysosomes Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/24be5d9c23dc42129117416062f59e64
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spelling	oai:doaj.org-article:24be5d9c23dc42129117416062f59e642021-11-15T15:22:02ZAn Intracellular Ammonium Transporter Is Necessary for Replication, Differentiation, and Resistance to Starvation and Osmotic Stress in <named-content content-type="genus-species">Trypanosoma cruzi</named-content>10.1128/mSphere.00377-172379-5042https://doaj.org/article/24be5d9c23dc42129117416062f59e642018-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00377-17https://doaj.org/toc/2379-5042ABSTRACT Trypanosoma cruzi, the etiologic agent of Chagas disease, undergoes drastic metabolic changes when it transits between a vector and mammalian hosts. Amino acid catabolism leads to the production of ammonium (NH4+), which needs to be detoxified. However, T. cruzi does not possess a urea cycle, and it is unknown how intracellular levels of ammonium are controlled. In this work, we identified an intracellular ammonium transporter of T. cruzi (TcAMT) that localizes to acidic compartments (reservosomes, lysosomes). TcAMT has 11 transmembrane domains and possesses all conserved and functionally important amino acid residues that form the pore in other ammonium transporters. Functional expression in Xenopus oocytes followed by a two-electrode voltage clamp showed an inward current that is NH4+ dependent at a resting membrane potential (Vh) lower than −120 mV and is not pH dependent, suggesting that TcAMT is not an NH4+/H+ cotransporter but an NH4+ or NH3/H+ transporter. Ablation of TcAMT by clustered regularly interspaced short palindromic repeat analysis with Cas9 (CRISPR-Cas9) resulted in significant defects in epimastigote and amastigote replication, differentiation, and resistance to starvation and osmotic stress. IMPORTANCE Trypanosoma cruzi is an important human and animal pathogen and the etiologic agent of Chagas disease. The parasite undergoes drastic changes in its metabolism during its life cycle. Amino acid consumption becomes important in the infective stages and leads to the production of ammonia (NH3), which needs to be detoxified. We report here the identification of an ammonium (NH4+) transporter that localizes to acidic compartments and is important for replication, differentiation, and resistance to starvation and osmotic stress.Teresa Cruz-BustosEvgeniy PotapenkoMelissa StoreyRoberto DocampoAmerican Society for MicrobiologyarticleTrypanosoma cruziamino acidsammoniaammoniumautophagylysosomesMicrobiologyQR1-502ENmSphere, Vol 3, Iss 1 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Trypanosoma cruzi amino acids ammonia ammonium autophagy lysosomes Microbiology QR1-502
spellingShingle	Trypanosoma cruzi amino acids ammonia ammonium autophagy lysosomes Microbiology QR1-502 Teresa Cruz-Bustos Evgeniy Potapenko Melissa Storey Roberto Docampo An Intracellular Ammonium Transporter Is Necessary for Replication, Differentiation, and Resistance to Starvation and Osmotic Stress in <named-content content-type="genus-species">Trypanosoma cruzi</named-content>
description	ABSTRACT Trypanosoma cruzi, the etiologic agent of Chagas disease, undergoes drastic metabolic changes when it transits between a vector and mammalian hosts. Amino acid catabolism leads to the production of ammonium (NH4+), which needs to be detoxified. However, T. cruzi does not possess a urea cycle, and it is unknown how intracellular levels of ammonium are controlled. In this work, we identified an intracellular ammonium transporter of T. cruzi (TcAMT) that localizes to acidic compartments (reservosomes, lysosomes). TcAMT has 11 transmembrane domains and possesses all conserved and functionally important amino acid residues that form the pore in other ammonium transporters. Functional expression in Xenopus oocytes followed by a two-electrode voltage clamp showed an inward current that is NH4+ dependent at a resting membrane potential (Vh) lower than −120 mV and is not pH dependent, suggesting that TcAMT is not an NH4+/H+ cotransporter but an NH4+ or NH3/H+ transporter. Ablation of TcAMT by clustered regularly interspaced short palindromic repeat analysis with Cas9 (CRISPR-Cas9) resulted in significant defects in epimastigote and amastigote replication, differentiation, and resistance to starvation and osmotic stress. IMPORTANCE Trypanosoma cruzi is an important human and animal pathogen and the etiologic agent of Chagas disease. The parasite undergoes drastic changes in its metabolism during its life cycle. Amino acid consumption becomes important in the infective stages and leads to the production of ammonia (NH3), which needs to be detoxified. We report here the identification of an ammonium (NH4+) transporter that localizes to acidic compartments and is important for replication, differentiation, and resistance to starvation and osmotic stress.
format	article
author	Teresa Cruz-Bustos Evgeniy Potapenko Melissa Storey Roberto Docampo
author_facet	Teresa Cruz-Bustos Evgeniy Potapenko Melissa Storey Roberto Docampo
author_sort	Teresa Cruz-Bustos
title	An Intracellular Ammonium Transporter Is Necessary for Replication, Differentiation, and Resistance to Starvation and Osmotic Stress in <named-content content-type="genus-species">Trypanosoma cruzi</named-content>
title_short	An Intracellular Ammonium Transporter Is Necessary for Replication, Differentiation, and Resistance to Starvation and Osmotic Stress in <named-content content-type="genus-species">Trypanosoma cruzi</named-content>
title_full	An Intracellular Ammonium Transporter Is Necessary for Replication, Differentiation, and Resistance to Starvation and Osmotic Stress in <named-content content-type="genus-species">Trypanosoma cruzi</named-content>
title_fullStr	An Intracellular Ammonium Transporter Is Necessary for Replication, Differentiation, and Resistance to Starvation and Osmotic Stress in <named-content content-type="genus-species">Trypanosoma cruzi</named-content>
title_full_unstemmed	An Intracellular Ammonium Transporter Is Necessary for Replication, Differentiation, and Resistance to Starvation and Osmotic Stress in <named-content content-type="genus-species">Trypanosoma cruzi</named-content>
title_sort	intracellular ammonium transporter is necessary for replication, differentiation, and resistance to starvation and osmotic stress in <named-content content-type="genus-species">trypanosoma cruzi</named-content>
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/24be5d9c23dc42129117416062f59e64
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An Intracellular Ammonium Transporter Is Necessary for Replication, Differentiation, and Resistance to Starvation and Osmotic Stress in <named-content content-type="genus-species">Trypanosoma cruzi</named-content>

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